The present invention relates to the use of self-assembled monolayers with mixtures of conductive oligomers and insulators to detect target analytes.
There are a number of assays and sensors for the detection of the presence and/or concentration of specific substances in fluids and gases. Many of these rely on specific ligand/antiligand reactions as the mechanism of detection. That is, pairs of substances (i.e. the binding pairs or ligand/antiligands) are known to bind to each other, while binding little or not at all to other substances. This has been the focus of a number of techniques that utilize these binding pairs for the detection of the complexes. These generally are done by labeling one component of the complex in some way, so as to make the entire complex detectable, using, for example, radioisotopes, fluorescent and other optically active molecules, enzymes, etc.
Other assays rely on electronic signals for detection, Of particular interest are biosensors. At least two types of biosensors are known; enzyme-based or metabolic biosensors and binding or bioaffinity sensors. See for example U.S. Pat. Nos. 4,713,347; 5,192,507; 4,920,047; 3,873,267; and references disclosed therein. While some of these known sensors use alternating current (AC) techniques, these techniques are generally limited to the detection of differences in bulk (or dielectric) impedance.
The use of self-assembled monolayers (SAMs) on surfaces for binding and detection of biological molecules has recently been explored. See for example WO98/20162; PCT US98/12430; PCT US98/12082; PCT US99/01705; and U.S. Pat. No. 5,620,850; and references cited therein.
Accordingly, it is an object of the invention to provide novel methods and compositions for the electronic detection of target analytes using self-assembled monolayers.
In accordance with the objects outlined above, the present invention provides compositions comprising electrodes comprising a monolayer comprising conductive oligomers, and a capture binding ligand. The composition further comprises a recruitment linker that comprises at least one covalently attached electron transfer moiety, and a solution binding ligand that will bind to a target analyte.
In a further embodiment, the invention provides methods of detecting a target analyte in a test sample comprising attaching said target analyte to an electrode comprising a monolayer of conductive oligomers via binding to a capture binding ligand. Recruitment linkers, or signal carriers, are directly or indirectly attached to the target analyte to form an assay complex. The method further comprises detecting electron transfer between said ETM and said electrode.
Kits and apparatus comprising the compositions of the method are also provided.